Antibody formulations

a technology of antibody and formulation, applied in the field of antibody formulation, can solve the problem that the liquid formulation is not suitable for lyophilization, and achieve the effect of reducing the reversible self-association

Inactive Publication Date: 2012-01-12
MEDIMMUNE LLC
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  • Summary
  • Abstract
  • Description
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AI Technical Summary

Benefits of technology

[0005]The present disclosure provides formulation methodology for reducing reversible self-association of an antibody or antibody fragment in a formulation. As such, the present disclosure provides methods that can be used to identify improved formulations for preparing antibodies and antibody fragments. Such formulations can be used, for example, therapeutically or diagnostically in a clinical or laboratory setting.
[0017]In certain embodiments of any of the foregoing aspects and embodiments, the formulation is suitable for intravenous administration. In certain embodiments, the aqueous carrier is water. In some embodiments, the formulation is non-pyrogenic. In other embodiments, the formulation further comprises a preservative to extend shelf-life. In some embodiments, the formulation is stable at 2-8° C. for at least 2 years as determined by high performance size exclusion chromatography (HPSEC). In some embodiments, purity of said formulation decreases by less than 0.5% per year for at least two years as determined by high performance size exclusion chromatography (HPSEC).
[0033]In further aspects, the present disclosure provides a method of eliminating or reducing reversible self-association (RSA) of an antibody in a formulation, comprising: providing an initial formulation comprising an antibody or antibody fragment, wherein RSA of said antibody or antibody fragment in said initial formulation is measurable by HPSEC i) at approximately 2-8° C. and / or ii) at a concentration of greater than 4 mg / ml, and which antibody or antibody fragment in said initial formulation contains high molecular weight forms; adding sucrose to said initial formulation to provide an altered formulation having about 4% to about 20% (weight / volume) sucrose, wherein RSA of the antibody or antibody fragment in the altered formulation is eliminated or reduced relative to that of the initial formulation, when compared at a given antibody concentration at approximately 2-8° C.
[0038]In other aspects, the present disclosure provides a method of eliminating or reducing reversible self-association (RSA) of an antibody in a formulation, comprising: a) providing an initial formulation comprising an antibody or antibody fragment, wherein RSA of said antibody or antibody fragment in said initial formulation is detectable by HPSEC at approximately 2-8° C. when assessed at a concentration of 10 mg / ml; b) assaying a biophysical property of said antibody or antibody fragment in said initial formulation using one or more assays; c) adjusting sucrose content of said initial formulation to achieve a final concentration from about 4% to about 20% (w / v) to produce an altered formulation; and d) assaying a biophysical property of said antibody or antibody fragment in said altered formulation using one or more assays; wherein RSA of the antibody or antibody fragment in the altered formulation is eliminated relative to that of the initial formulation, when compared at a given antibody concentration at approximately 2-8° C.

Problems solved by technology

In one embodiment, the liquid formulation is not suitable for lyophilization.

Method used

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  • Antibody formulations
  • Antibody formulations
  • Antibody formulations

Examples

Experimental program
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Effect test

example 1

Initial Characterization of the RSA Tendency of MabA in an Initial Formulation—RSA of MabA is Dependent on Temperature and / or Concentration

[0242]RSA tendency of MabA in an initial, non-sucrose containing formulation was evaluated at an antibody concentration of 10 mg / ml. In addition to antibody, the initial formulation contained, in water, 50 mM sodium-acetate, 100 mM NaCl, 0.01% (w / v) PS80, pH 5.5. At 2-8° C. and at a protein concentration of 10 mg / ml, a shoulder on the leading edge of the monomer peak was detected by HPSEC. The HPSEC shoulder was not observed after equilibration of MabA for about 80 minutes at 23-27° C. or following dilution to a protein concentration of about 4 mg / ml. Characterization studies, including analytical ultracentrifugation (AUC), were performed to further understand the nature of the HPSEC shoulder of MabA. The results are summarized below.

[0243]The size distribution results obtained by AUC are consistent with the HPSEC data showing that shoulder forma...

example 2

Elimination of MabA RSA Tendency in Sucrose-Containing Formulation as Determined by HPSEC, AUC and DLS

MabA Exhibits RSA Tendency in Various Formulations Including the Acetate / Salt Formulation Under Cold Temperature Storage Conditions

[0250]The RSA tendency of MabA in an initial, non-sucrose-containing, acetate / salt formulation (10 mg / ml MabA, 50 mM Na-acetate, 100 mM NaC1, 0.01% (w / v) PS80, pH 5.5) was evaluated by high performance size exclusion chromatography (HPSEC) at standard storage temperature (at about 2-8° C.) and at room temperature (at about 23-27° C.). As shown in FIG. 1A, RSA tendency of MabA is pronounced at about 2-8° C., as indicated by a leading edge shoulder on the monomer peak. Such a leading edge shoulder was not detected when tested at 25° C., indicating that MabA behaves differently at 2-8° C. as compared at 23-27° C. More specifically, MabA exhibits significant RSA tendency at 2-8° C. that is measurable by HPSEC.

[0251]This result regarding the RSA characteristi...

example 3

RSA of MabA is Driven by Hydrophobic Interactions and is Correlated to its Hydrodynamic Radius

[0254]RSA is enhanced, in part, “by the exposure of hydrophobic surfaces and the ensuing hydrophobic intermolecular interactions. An evaluation of the interaction between a fluorescent dye (1-anilino-8-naphthalene sulfonate; “ANS”) with MabA illustrates this. ANS binds preferentially to hydrophobic patches on a protein surface, which results in an increase in ANS fluorescence intensity. Thus, a decrease in ANS fluorescence intensity is indicative of a decrease in available hydrophobic surfaces and correlates to a decrease in hydrodynamic radius.

[0255]As demonstrated in FIG. 3, the interactions between ANS and hydrophobic surfaces on MabA diminish when assessed in formulations having increased sucrose content. This is consistent with the conclusion that the antibody becomes progressively more compact when present in formulations having increased sucrose concentration, as assessed at 5° C. (e...

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Abstract

The present invention relates to formulations comprising sucrose, and methods of making such formulations, wherein the sucrose content promotes the reduction or elimination of the reversible self-association (RSA) tendency of the antibody in the formulation. The present invention also relates to formulations comprising an anti-PDGFR-alpha antibody or antibody fragment. Such antibodies can be used in various methods of treatment. The application further relates to a method of eliminating or reducing the RSA tendency of antibodies in a formulation.

Description

RELATED APPLICATIONS[0001]This application claims the benefit of U.S. provisional application 61 / 361,209, filed Jul. 2, 2010. The disclosure of the foregoing application is hereby incorporated by reference in its entirety.SEQUENCE LISTING [0002]The instant application contains a Sequence Listing which has been concurrently submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Jun. 24, 2011, is named 12728101.txt and is 3,275 bytes in size.BACKGROUND OF THE INVENTION[0003]Instability and / or presence of undesirable species in an antibody formulation may pose problems including simple inconvenience, such as the need to store or handle the formulation in a particular way, and safety and efficacy concerns, such as those caused by increased toxicity or immunogenicity. One characteristic that is observed with some antibodies is the tendency for the antibody molecules to self-associate reversibly, also known as reversible ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/395A61P35/00
CPCA61K9/0019A61K47/26C07K16/00C07K16/2863A61K39/39591A61K47/12A61P35/00
Inventor DIMITROVA, MARIANA N.MODY, NEIL
Owner MEDIMMUNE LLC
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